A Novel Construct of Coral Granules-Poly-L-Lactic Acid Nanomembrane Sandwich Double Stem Cell Sheet Transplantation as Regenerative Therapy of Bone Defect Model.

OBJECTIVES

We examined the use of a new approach in nanotechnology and stem cell research as regenerative therapy for bone tissue defects.

MATERIALS AND METHODS

We compared in vitro osteogenic potential of human Wharton jelly mesenchymal stem cells using coral granules and poly-L-lactic acid nanofiber according to proliferation (by cck-8 kit) and osteogenes (runt-related transcription factor 2, alkaline phosphatase, osteonectin) by quantitative reverse transcription-polymerase chain reaction, alkaline phosphatase assay, calcium measurement, and assessment of mineralization by Alizarin red and von Kossa staining. To overcome the limitations of natural coral, we made a modification by packaging the coral granules-human Wharton jelly mesenchymal stem cells by nanomembrane-human Wharton jelly mesenchymal stem cells to form sandwich double cell sheets and compared this hole with other holes (one was filled by human Wharton jelly mesenchymal stem cell suspension, and the other was filled by coral granules saturated with preinduced mesenchymal stem cells) by radiological and histopathological studies for repairing the bone gap.

RESULTS

Collagen-coated poly-L-lactic acid showed higher mRNA levels for all osteogenes (P < .001), higher alkaline phosphatase and calcium content (P < .001), and greater stainability. Our in vivo experiment showed that the holes implanted with sandwich double cell sheet-poly-L-lactic acid coral were completely filled mature compact bone. The holes implanted with human Wharton jelly mesenchymal stem cells alone were filled with immature compact bone. Holes implanted with coral granules-human Wharton jelly mesenchymal stem cells were filled with condensed connective tissue.

CONCLUSIONS

Poly-L-lactic acid nanofiber has greater osteogenic differentiating effect than the coral granules. The new approach of sDCS-PLLA-coral construct proved success for bone regeneration and repairing the bone gap and this may improve the design of tissue constructs for bone tissue regenerative therapy.